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Visualisation of cerebrospinal fluid flow patterns in albino Xenopus larvae in vivo

机译：白化非洲爪蟾幼虫体内脑脊液流动模式的可视化

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Background It has long been known that cerebrospinal fluid (CSF), its composition and flow, play an important part in normal brain development, and ependymal cell ciliary beating as a possible driver of CSF flow has previously been studied in mammalian fetuses in vitro. Lower vertebrate animals are potential models for analysis of CSF flow during development because they are oviparous. Albino Xenopus laevis larvae are nearly transparent and have a straight, translucent brain that facilitates the observation of fluid flow within the ventricles. The aim of these experiments was to study CSF flow and circulation in vivo in the developing brain of living embryos, larvae and tadpoles of Xenopus laevis using a microinjection technique. Methods The development of Xenopus larval brain ventricles and the patterns of CSF flow were visualised after injection of quantum dot nanocrystals and polystyrene beads (3.1 or 5.8 μm in diameter) into the fourth cerebral ventricle at embryonic/larval stages 30-53. Results The fluorescent nanocrystals showed the normal development of the cerebral ventricles from embryonic/larval stages 38 to 53. The polystyrene beads injected into stage 47-49 larvae revealed three CSF flow patterns, left-handed, right-handed and non-biased, in movement of the beads into the third ventricle from the cerebral aqueduct (aqueduct of Sylvius). In the lateral ventricles, anterior to the third ventricle, CSF flow moved anteriorly along the outer wall of the ventricle to the inner wall and then posteriorly, creating a semicircle. In the cerebral aqueduct, connecting the third and fourth cerebral ventricles, CSF flow moved rostrally in the dorsal region and caudally in the ventral region. Also in the fourth ventricle, clear dorso-ventral differences in fluid flow pattern were observed. Conclusions This is the first visualisation of the orchestrated CSF flow pattern in developing vertebrates using a live animal imaging approach. CSF flow in Xenopus albino larvae showed a largely consistent pattern, with the exception of individual differences in left-right asymmetrical flow in the third ventricle.
机译：背景技术早就知道，脑脊液（CSF），其组成和流动在正常的大脑发育中起着重要的作用，并且以前曾在体外对哺乳动物胎儿进行过研究，室管细胞纤毛跳动是CSF流动的可能驱动因素。由于它们是卵生的，低等脊椎动物是发育过程中CSF流量分析的潜在模型。白化非洲爪蟾幼虫几乎是透明的，并具有笔直的半透明的大脑，有助于观察脑室内的流体流动。这些实验的目的是使用显微注射技术研究非洲爪蟾活体胚胎，幼虫和t发育中大脑中的脑脊液流动和循环。方法在胚胎/幼虫阶段30-53向第四脑室注射量子点纳米晶体和聚苯乙烯珠（直径为3.1或5.8μm）后，观察非洲爪蟾幼虫脑室的发育和CSF流动模式。结果荧光纳米晶体显示出从胚胎/幼虫阶段38到53阶段脑室的正常发育。在47-49阶段幼虫中注射的聚苯乙烯珠粒显示了三种CSF流动模式，左旋，右旋和无偏。珠子从脑导水管（西尔维斯导水管）进入第三脑室。在第三脑室之前的侧脑室中，脑脊液流沿脑室的外壁向前移动至内壁，然后向后移动，从而形成半圆。在连接第三脑室和第四脑室的脑导水管中，脑脊液流在背侧区域向后向运动，在腹侧区域向后向运动。同样在第四脑室，观察到明显的背-腹液流模式差异。结论这是使用活体动物成像方法对发育中的脊椎动物中精心策划的CSF流动模式进行的首次可视化。非洲爪蟾白化幼虫的CSF流量显示出基本一致的模式，但第三脑室的左右不对称流量存在个体差异。

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